Background: Crown-like structures in breast adipose tissue (CLS-B), composed of necrotic adipocytes encircled by macrophages, are associated with obesity and hypothesized to worsen breast cancer prognosis; however, data are sparse, particularly in multi-racial populations. Methods:We assessed specimens for CLS-B from 174 African-American and 168 White women with stage I-III breast cancer treated by mastectomy. Benign breast tissue from an uninvolved quadrant was immunohistochemically stained for CD68 to determine CLS-B presence and density (per cm 2 of adipose tissue). Demographic and lifestyle factors, collected via medical record review, were analyzed for associations with CLS-B using logistic regression. Multivariable Cox proportional hazards models were used to compute hazard ratios (HRs) and 95% confidence intervals (CIs) for associations between CLS-B and overall (OS) or progression-free (PFS) survival.Results: Detection of any CLS-B was similar between African-American (32%) and White (29%) patients with no evidence of an association between race and CLS-B in multivariable models (OR = 0.82, 95% CI = 0.49-1.36). Detection of CLS-B was associated with obesity (OR = 4.73, 95% CI = 2.48-9.01) and age ≥ 60 years at diagnosis (OR = 1.78, 95% CI = 0.99-3.21). There was some evidence of associations with parity and current smoking status. Detection of CLS-B was not associated with OS (HR = 1.02, 95% CI = 0.55-1.87) or PFS (HR = 0.99, 95% CI = 0.59-1.67). Conclusions: Our results show a strong, positive association between BMI and CLS-B in non-tumor tissue similar to previous findings. Detection of CLS-B did not vary by race and was not associated with worse OS or PFS.

We have previously shown that selective inhibition of histone deacetylase 3 (HDAC3) decreases infarct volume and improves long-term functional outcomes after stroke. In this study, we examined the effects of HDAC3 inhibition on cerebral edema and blood–brain barrier (BBB) leakage and explored its underlying mechanisms. Adult male Wistar rats were subjected to 2-h middle cerebral artery occlusion (MCAO) and randomly treated i.p. with either vehicle or a selective HDAC3 inhibitor (RGFP966) at 2 and 24 h after stroke. Modified neurological severity scores (mNSS) were calculated at 2 h, 1 day, and 3 days. H&E, Evans blue dye (EBD) assay, and fluorescein isothiocyanate (FITC)-dextran were employed to assess cerebral edema and BBB leakage. Western blot for matrix metalloproteinase-9 (MMP9), MMP-9 zymography, and immunostaining for HDAC3, GFAP, Iba-1, albumin, aquaporin-4, claudin-5, ZO-1, and NF-kB were performed. Early RGFP966 administration decreased cerebral edema (p = 0.002) and BBB leakage, as measured by EBD assay, FITC-dextran, and albumin extravasation (p < 0.01). RGFP966 significantly increased tight junction proteins (claudin-5 and ZO-1) in the peri-infarct area. RGFP966 also significantly decreased HDAC3 in GFAP + astrocytes, which correlated with better mNSS (r = 0.67, p = 0.03) and decreased cerebral edema (r = 0.64, p = 0.04). RGFP966 decreased aquaporin-4 in GFAP + astrocytes (p = 0.002), as well as, the inflammatory markers Iba-1, NF-kB, and MMP9 in the ischemic brain (p < 0.05). Early HDAC3 inhibition decreases cerebral edema and BBB leakage. BBB protection by RGFP966 is mediated in part by the upregulation of tight junction proteins, downregulation of aquaporin-4 and HDAC3 in astrocytes, and decreased neuroinflammation.

Background: Crown-like structures in breast adipose tissue (CLS-B), indicative of proinflammatory conditions, are most frequently observed among obese (body mass index, BMI ≥30 kg/m2) women and may contribute to poor prognosis in this group. African-American (AA) women have disproportionately higher rates of obesity than White women, and at least one prior study suggests the prevalence of CLS-B may be higher among AA women. However, most previous studies have examined CLS-B within affected tissues, which may reflect inflammation in the tumor microenvironment, and few have examined the association between CLS-B and clinical outcomes by race. Methods: We examined the presence of CLS-B detected by CD68 immunohistochemistry in normal adjacent breast tissue from a quadrant uninvolved by tumor obtained via mastectomy among 174 African-American women and 168 White women with stage I—III breast cancer diagnosed at Emory University Hospitals (2007—2012). We also investigated associations between CLS-B and other demographic and lifestyle factors at diagnosis (e.g., BMI, smoking status, age at menarche, parity, lactation, menopausal status, hormone replacement therapy use, and family history of breast cancer). Patients were followed for an average of seven years after diagnosis for recurrence and survival. Multivariable Cox proportional hazards models were used to compute hazard ratios (HR) and 95% confidence intervals (CI) for associations between CLS-B presence and progression-free survival (PFS), controlling for BMI and other potential confounders. Results: Median age at diagnosis for both AA and White women was 54 years, with more than 60% postmenopausal among both groups. AA women were more likely than White women to be obese (52% vs. 24%) and have ER- tumors (30% vs. 12%). Presence of any CLS-B was similar between AA (32%) and White (29%) patients. In multivariable models, we did not find any association between CLS-B and race (HR=1.14, 95% CI: 0.72, 1.82) with the only statistically significant factors being BMI (≥30 vs. 18.5-&lt;25 kg/m2: HR=4.36, 95% CI: 2.17, 8.76) and parity (1+ vs. 0 births: HR=0.43, 95% CI: 0.21, 0.91). Over follow-up, 46 breast cancer recurrences and 52 deaths (23 from breast cancer) occurred. Overall, the presence of CLS-B was not associated with PFS (multivariable HR: 0.97, 95% CI: 0.58, 1.62). When examined by race, there was a difference in the direction of the association between CLS-B and PFS among AA women (HR=1.25, 95% CI: 0.64, 2.46) compared to White women (HR=0.75, 95% CI: 0.33, 1.71), although this difference was not statistically significant (P=0.86). Conclusion: Our results show a strong, positive association between BMI and CLS-B in non-tumor tissue and an inverse association with parity. We did not observe a difference in CLS-B presence by race nor did we find CLS-B to be associated with worse progression-free survival, which is in contrast to previous studies that have examined the presence of CLS-B within specimens in close proximity to the tumor. Citation Format: Aswathy M Cheriyan, Mark E Sherman, Yuan Liu, Keerthi Gogineni, Jiaqi Liu, Jiabei He, Uma Krishnamurti, Ryan Ashiqueali, Jinjing He, Rami Yacoub, Jasmine Miller-Kleinhenz, Lauren E McCullough, Maret L Maliniak. Presence of crown-like structures in breast adipose tissue and clinical outcomes among African-American and White breast cancer patients [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr C074.